Short-term organoid culture for drug sensitivity testing of high-grade serous carcinoma.
Short-term organoid culture for drug sensitivity testing of high-grade serous carcinoma.
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DOI：
10.1016/j.ygyno.2020.03.026
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发表时间：
2020-06
影响因子：
4.7
通讯作者：
Smith LH
中科院分区：
文献类型：
--
作者：
Chen H;Gotimer K;De Souza C;Tepper CG;Karnezis AN;Leiserowitz GS;Chien J;Smith LH
癌症患者来源的类器官（PDO）在细胞外基质的存在下生长为三维（3D）结构，并且已经发现代表原始肿瘤的遗传复杂性。此外，PDO可以在较短的时间内生长并进行药物敏感性测试，并且比患者来源的异种移植模型花费更少。许多复发性卵巢癌患者出现恶性积液，化疗难以治愈。由于这些相同的患者经常出现姑息性腹水或胸腔积液抽吸，因此有可能获得恶性渗出液中存在的多细胞球体（MCS）形式的肿瘤标本。我们的目标是在选择支持类器官生长的条件下，从卵巢癌恶性积液中培养MCS，并将其用作经验性药物敏感性测试的平台。在这项研究中，恶性积液标本收集自高级别浆液性卵巢癌（HGSOC）患者。回收多组分灭菌剂，并使其经受旨在支持类器官生长的培养条件。在一个样本子集中，在短期培养期间的两个时间点进行RNA测序，以确定转录组响应于培养条件的变化。还使用Ki 67染色和组织学分析在这些标本中表征类器官诱导。对所有标本进行药物敏感性试验。我们的模型描述了在原代培养数天内形成的类器官，其可以概括恶性腹水的组织学特征，并且可以扩增至少6天。对4例患者标本的RNA-seq分析显示，在培养的6天内，与细胞增殖、上皮-间充质转化和KRAS信号通路相关的基因显著上调。药物敏感性试验确定了几种具有治疗潜力的药物。来自HGSOC恶性积液的MCS的短期类器官培养可用作经验性药物敏感性测试的平台。这些离体模型可能有助于在个体化治疗选择之前筛选新的或现有的治疗剂。
Cancer patient-derived organoids (PDOs) grow as three dimensional (3D) structures in the presence of extracellular matrix and have been found to represent the original tumor’s genetic complexity. In addition, PDOs can be grown and subjected to drug sensitivity testing in a shorter time course and with lesser expense than patient-derived xenograft models. Many patients with recurrent ovarian cancer develop malignant effusions that become refractory to chemotherapy. Since these same patients often present for palliative aspiration of ascites or pleural effusions, there is a potential opportunity to obtain tumor specimens in the form of multicellular spheroids (MCS) present in malignant effusion fluids. Our objective was to develop a short duration culture of MCS from ovarian cancer malignant effusions in conditions selected to support organoid growth and use them as a platform for empirical drug sensitivity testing. In this study, malignant effusion specimens were collected from patients with high-grade serous ovarian carcinoma (HGSOC). MCS were recovered and subjected to culture conditions designed to support organoid growth. In a subset of specimens, RNA-sequencing was performed at two time points during the short-term culture to determine changes in transcriptome in response to culture conditions. Organoid induction was also characterized in these specimens using Ki67 staining and histologic analysis. Drug sensitivity testing was performed on all specimens. Our model describes organoids formed within days of primary culture, which can recapitulate the histological features of malignant ascites fluid and can be expanded for at least 6 days. RNA-seq analysis of four patient specimens showed that within 6 days of culture, there was significant up-regulation of genes related to cellular proliferation, epithelial-mesenchymal transition, and KRAS signaling pathways. Drug sensitivity testing identified several agents with therapeutic potential. Short duration organoid culture of MCS from HGSOC malignant effusions can be used as a platform for empiric drug sensitivity testing. These ex vivo models may be helpful in screening new or existing therapeutic agents prior to individualized treatment options.
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DOI：
10.1097/igc.0000000000001061
发表时间：
2017-10
期刊：
International journal of gynecological cancer : official journal of the International Gynecological Cancer Society
影响因子：
--
作者：
Girda E;Huang EC;Leiserowitz GS;Smith LH
通讯作者：
Smith LH
影响因子：
28.2
作者：
Hill SJ;Decker B;Roberts EA;Horowitz NS;Muto MG;Worley MJ Jr;Feltmate CM;Nucci MR;Swisher EM;Nguyen H;Yang C;Morizane R;Kochupurakkal BS;Do KT;Konstantinopoulos PA;Liu JF;Bonventre JV;Matulonis UA;Shapiro GI;Berkowitz RS;Crum CP;D'Andrea AD
通讯作者：
D'Andrea AD
影响因子：
9.9
作者：
Jabs J;Zickgraf FM;Park J;Wagner S;Jiang X;Jechow K;Kleinheinz K;Toprak UH;Schneider MA;Meister M;Spaich S;Sütterlin M;Schlesner M;Trumpp A;Sprick M;Eils R;Conrad C
通讯作者：
Conrad C
影响因子：
45.3
作者：
EINZIG, AI;WIERNIK, PH;GOLDBERG, GL
通讯作者：
GOLDBERG, GL
影响因子：
8
作者：
Bykov, VJN;Zache, N;Wiman, KG
通讯作者：
Wiman, KG